Ectopic hCG-beta secreted by DoT and CaSki human carcinoma cell lines contains two components, ectopic hCG-beta-I and ectopic hCG-beta-II. Ectopic hCG-beta-II lacks the C-terminal peptide (CTP), based on lack of recognition by antibodies against CTP, lack of susceptibility to thermolysin (which normally cleaves the CTP), and lack of binding of the desialylated derivative to Arachis hypogaea (AH) lectin (specific for Gal-GalNAc of the O-linked oligosaccharides on the CTP). Immunoreactive hCG-beta-like material has been detected in the serum and urine of two of four patients with cervical carcinoma. Conditions for large-scale roller bottle culture of CaSki cervical carcinoma cells were optimized. A method to separate ectopic hCG-beta-I from ectopic hCG-beta-II has been validated and involves differential affinity chromatography with: (1)\anti-hCG-alpha Sepharose; (2)\anti-[CTP-directed]hCG-beta-Sepharose; and (3)\anti-[core-directed]hCG-beta Sepharose. Ectopic hCG-beta-II is eluted from the third stage with 4 M guanidine\HC1, pH 3, and desalted. The purified ectopic hCG-beta-II failed to combine with standard hCG-alpha under conditions where standard hCG-beta combined with standard hCG-alpha. Evidence was obtained for the presence of additional carbohydrates on ectopic hCG-beta. (C)